LOCUS       sequence-244740-        7935 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses mouse PCSK7 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7935)
  AUTHORS   Li JJ, Liu J, Lupino K, Liu X, Zhang L, Pei L
  TITLE     Growth Differentiation Factor 15 Maturation Requires Proteolytic 
            Cleavage by PCSK3, -5, and -6.
  JOURNAL   Mol Cell Biol. 2018 Oct 15;38(21). pii: MCB.00249-18. doi: 
            10.1128/MCB.00249-18. Print 2018 Nov 1.
  PUBMED    30104250
REFERENCE   2  (bases 1 to 7935)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell 
            Biol."; date: "2018-10-15"; volume: "38(21). pii: MCB.00249-18. 
            doi"; pages: " 10.1128/MCB.00249-18. Print 2018 Nov 1"
COMMENT     Sequence Label: pcDNA3.1-mPCSK7
FEATURES             Location/Qualifiers
     source          1..7935
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3433..3474
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3484..3501
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3524..3541)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3530..3754
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3800..4228
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3887..3906)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4097..4118
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4237..4257)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4242..4571
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4422..4557
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4484..4503
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4638..5432
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4692..4711)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5302..5321
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5608..5729
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5645..5664)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5699..5718
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5778..5794)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5778..5794)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5791..5813)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5802..5818
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5826..5832,5833..5850,5851..5856))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5871..5892
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6009..6026)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6180..6768)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6260..6279)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6939..7730,7731..7799))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7562..7581
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7800..7904)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttggt accgagctcg gatccactag tccagtgtgg tggaattgcc cttagggcaa
      961 ttctgcagat atctcacaag atgagtgtgg tatggttacc caggccagga cctaaacttc
     1021 acagagagcc cagcctgctg ttctgatgcc gaaagggagg cagaaagtcc cacacttgga
     1081 tgcccacctg ggcctgccca tctgcctctg gctggaatta gccatcttct ttctggttcc
     1141 ccaggtcatg ggcctatcag aggcaggtgg gcttgacatc ttgggcacag gggggctgag
     1201 ctgggccgta catctggaca gcctagaagg tgagaggaag gaagagagtc tgacgcaaca
     1261 ggccgatgct gtggcccagg cagcagggct tgtgaatgct gggcgcattg gagaactcca
     1321 ggggcactac ctctttgtcc agcctactgg gcataggcaa gccatggagg tggaggccat
     1381 gcggcaacag gcagaggctg tgttagccag gcatgaagct gtgcgctggc actcggagca
     1441 aacgctgctg aagagggcca agcgcagtat ccacttcaat gatcccaagt accctcaaca
     1501 gtggcacctg aataatcggc ggagcccagg aagagacatc aatgtgacag gtgtgtggga
     1561 gcgaaatgta actgggcgtg gggtgacagt ggtggtggtg gatgacggag tggaacacac
     1621 cgtccaggac atcgcaccca actacagtcc agagggaagt tatgacctca actctaatga
     1681 cccagatccc atgccccacc ctgatgagga gaatggtaac caccatggga cccggtgtgc
     1741 aggagaaatt gcagctgtgc ccaacaacag cttctgtgca gtgggtgtgg cctacgggag
     1801 ccgaatagca ggtatccggg tgctggatgg accacttact gacagtatgg aggctgtggc
     1861 gttcaacaag cactatcaga tcaatgacat ctacagctgc agctggggcc cagatgatga
     1921 tgggaagaca gtggatggtc ctcatcagct cggaaaggct gccttacaac atggagtgat
     1981 ggctggtcgc caaggctttg gaagtatctt tgtggttgcc agtggtaatg ggggccagca
     2041 caatgacaac tgcaactatg atggctatgc caactccatc tacactgtca ccataggagc
     2101 tgtggatgag gagggacgga tgccttttta tgcagaggag tgtgcctcca tgctggcagt
     2161 caccttcagt ggtggagaca agatgctgcg gagcattgtg accactgact gggaccttca
     2221 gaagggcact ggctgcactg aaggccacac aggaacctca gctgcagctc ctctggcagc
     2281 tggaatgata gccctcatgc tgcaggtgcg gccctgcctc acatggcggg atgtacagca
     2341 cataattgtc ttcacagcca tccagtatga agatcatcat gcagactggc tcaccaatga
     2401 ggctggcttc agccacagcc atcagcatgg tttcggcctg ctcaacgctt ggagacttgt
     2461 caatgcagcc aagatctgga catctgtccc ttacttagcc tcctatgtca gccccatgct
     2521 gaaagaaaat aaggctgttc cgaggtcccc ccactctctg gaggtgctat ggaatgtcag
     2581 caggacggac ctggagatgt cggggctgaa gaccttggaa catgtggcag tgacagtctc
     2641 catcactcac ccacgacgtg gcagcttaga actgaaactg ttttgtccca gtggcatgat
     2701 gtctttgatc ggcgcgcccc gcagcatgga ctcggatcct aatggcttca atgcttggac
     2761 cttttccact gtgcggtgct ggggggaaag agcaagaggc gtctacagac tggttatcag
     2821 ggatgtagga gatgagccgc tccagatggg catcctccag cagtggcagc tgaccctgta
     2881 tggctccatg tggagtccag tagacatcaa ggacagacaa agtctcttag aaagtgctat
     2941 gagtggaaaa tacctgcatg atggcttcac cctgccttgc ccacctgggc tgaaaatccc
     3001 tgaggaggat ggctacacca ttacccctaa cacactcaag acccttgtgc tggtaggctg
     3061 cttcagtgtc ttctggacta tttactacat gctagaagtg tgcctgagcc agaggaacaa
     3121 ggcttccacc catggctgca ggaagggatg ctgcccctgg gccccacgaa ggcaaaactc
     3181 caaggatgca gggacagcac tagaatcaat gccactgtgc agcagcaagg acctggatgg
     3241 agtggattca gagcatgggg actgcacaac tgcatctagc ttcctggccc cagagctgga
     3301 ttgtcctcca caccaacccc cagacctgct gcaggggaag agcgggcaga tttgctgacc
     3361 ccagagccca gcttccccca tgtacaacag gcacttccta aactctcgag tctagagggc
     3421 ccgcggttcg aaggtaagcc tatccctaac cctctcctcg gtctcgattc tacgcgtacc
     3481 ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac tgtgccttct
     3541 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     3601 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     3661 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     3721 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     3781 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3841 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3901 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3961 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     4021 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4081 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     4141 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     4201 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     4261 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4321 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4381 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4441 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4501 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4561 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     4621 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4681 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4741 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4801 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4861 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4921 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4981 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     5041 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     5101 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     5161 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     5221 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     5281 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     5341 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     5401 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc
     5461 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     5521 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     5581 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     5641 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     5701 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     5761 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     5821 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     5881 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5941 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     6001 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     6061 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     6121 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     6181 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     6241 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     6301 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     6361 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     6421 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     6481 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     6541 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6601 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6661 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6721 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6781 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6841 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6901 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6961 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     7021 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     7081 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     7141 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     7201 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     7261 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     7321 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     7381 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     7441 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     7501 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     7561 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     7621 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     7681 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     7741 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     7801 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     7861 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     7921 gtgccacctg acgtc
//
